Department of Molecular Pathology, Nara Medical University, Kashihara, Japan.
Urol Oncol. 2012 May-Jun;30(3):306-13. doi: 10.1016/j.urolonc.2010.05.001. Epub 2010 Sep 16.
The colorimetric in situ hybridization (CISH)-based matrix metalloproteinase (MMP)-to-E-cadherin (ECD) ratio (MER) has been revealed as an excellent marker for the disease stage in prostate cancer. The one aim of this study was investigating a new method for estimation of MER by bicolor fluorescent ISH (bicolor FISH) with a computerized fluorescence detector-based system. Another aim was examination of relation of MER by bicolor FISH with expression of vascular endothelial growth factor-C (VEGF-C).
The bicolor FISH technique used cyanin 5 (cy5)-labeled MMP-2 and -9 probes, and a cyanin 3 (cy3)-labeled ECD probe on needle biopsy specimens from 67 prostate cancer cases. The ISH was followed by computerized detection of the signal intensities and cy5-to-cy3 ratios using a fluorescence detector. VEGF-C expression was examined using cy5-labeled VEGF-C by computerized detection.
The bicolor FISH-based MER was well correlated with CISH-based MER (P < 0.0001). The bicolor FISH-based MER correlated with Gleason score and pathologic stage of the cases. VEGF-C mRNA expression was associated with the pathologic stage and maximum lymph vessel density (LVD). The LVD was associated with VEGF-C expression at the tumor area where the maximum MER was detected (P < 0.0001).
The MER was correlated with the VEGF-C expression and LVD, indicating lymph node metastasis of prostate cancer. Therefore, this computer-assisted MER is a useful marker for preoperative prediction of disease stage, especially lymph node metastasis, of prostate cancer.
基于比色原位杂交(CISH)的基质金属蛋白酶(MMP)与上皮钙黏蛋白(ECD)比值(MER)已被证实为前列腺癌疾病分期的优秀标志物。本研究的目的之一是研究一种新的方法,通过基于计算机化荧光检测的双色荧光原位杂交(bicolor FISH)来估计 MER。另一个目的是研究 bicolor FISH 法测定的 MER 与血管内皮生长因子-C(VEGF-C)表达的关系。
该双色 FISH 技术使用花青 5(cy5)标记的 MMP-2 和 -9 探针,以及针吸活检标本中的花青 3(cy3)标记的 ECD 探针,对 67 例前列腺癌病例进行检测。ISH 后,使用荧光探测器对信号强度和 cy5 与 cy3 的比值进行计算机化检测。使用 cy5 标记的 VEGF-C 通过计算机化检测来检测 VEGF-C 的表达。
基于 bicolor FISH 的 MER 与基于 CISH 的 MER 高度相关(P < 0.0001)。基于 bicolor FISH 的 MER 与 Gleason 评分和病例的病理分期相关。VEGF-CmRNA 表达与病理分期和最大淋巴管密度(LVD)相关。在检测到最大 MER 的肿瘤区域,LVD 与 VEGF-C 表达相关(P < 0.0001)。
MER 与 VEGF-C 表达和 LVD 相关,表明前列腺癌的淋巴结转移。因此,这种计算机辅助的 MER 是预测前列腺癌术前疾病分期,特别是淋巴结转移的有用标志物。